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Visualizing siRNA in Mammalian Cells: Fluorescence Analysis of the RNAi Effect

Olympus BX60 microscope with the appropriate fluorescence filters. Photomicroscopy was performed using a Hitachi KP-C571 camera.


RNA Isolation and Analysis
Total RNA from cell cultures was extracted using the RNAqueous-4PCR Kit (Ambion) and quantitated by spectrophotometer. The expression level of both target and control genes was determined for each experimental sample using the NorthernMax-Gly Kit (Ambion).


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The Silencer siRNA Labeling Kits contain reagents manufactured for Ambion by MIRUS.

References

  1. Brown D, Jarvis R, Pallotta V, Byrom M, Ford L (2002) RNA Interference in Mammalian Cell Culture: Design, Execution and Analysis of the siRNA effect. Ambion TechNotes 9(1): 3-5.
  2. Demeterco C, Itkin-Ansari P, Tyrberg B, Ford LP, Jarvis RA, Levine F (2002) c-Myc controls proliferation versus differentiation in human pancreatic endocrine cells. JCEM (in press).
  3. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, and Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411: 494-498.
  4. Hammond SM, Boettcher S, Caudy AA, Kobayashi R, and Hannon GJ (2001) Argonaute2, a link between genetic and
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